Clinical application of a systems model of apoptosis execution for the prediction of colorectal cancer therapy responses and personalisation of therapy

Objective Key to the clinical management of colorectal cancer is identifying tools which aid in assessing patient prognosis and determining more effective and personalised treatment strategies. We evaluated whether an experimental systems biology strategy which analyses the susceptibility of cancer cells to undergo caspase activation can be exploited to predict patient responses to 5-fluorouracil-based chemotherapy and to case-specifically identify potential alternative targeted treatments to reactivate apoptosis. Design We quantified five essential apoptosis-regulating proteins (Pro-Caspases 3 and 9, APAF-1, SMAC and XIAP) in samples of Stage II (n=13) and III (n=17) tumour and normal colonic (n=8) tissue using absolute quantitative immunoblotting and employed systems simulations of apoptosis signalling to predict the susceptibility of tumour cells to execute apoptosis. Additional systems analyses assessed the efficacy of novel apoptosis-inducing therapeutics such as XIAP antagonists, proteasome inhibitors and Pro-Caspase-3-activating compounds in restoring apoptosis execution in apoptosis-incompetent tumours. Results Comparisons of caspase activity profiles demonstrated that the likelihood of colorectal tumours to undergo apoptosis decreases with advancing disease stage. Systems-level analysis correctly predicted positive or negative outcome in 85% (p=0.004) of colorectal cancer patients receiving 5-fluorouracil based chemotherapy and significantly outperformed common uni- and multi-variate statistical approaches. Modelling of individual patient responses to novel apoptosis-inducing therapeutics revealed markedly different inter-individual responses. Conclusions Our study represents the first proof-of-concept example demonstrating the significant clinical potential of systems biology-based approaches for predicting patient outcome and responsiveness to novel targeted treatment paradigms.

[1]  M. Morita,et al.  Clinical significance of Smac/DIABLO expression in colorectal cancer. , 2009, Oncology reports.

[2]  Andrew J. Wilson,et al.  Molecular mechanisms of action and prediction of response to oxaliplatin in colorectal cancer cells , 2004, British Journal of Cancer.

[3]  P. Engstrom,et al.  Phase I Study of Capecitabine and Oxaliplatin in Combination With the Proteasome Inhibitor Bortezomib in Patients With Advanced Solid Tumors , 2008, American journal of clinical oncology.

[4]  G. Giaccone,et al.  Expression of X-linked inhibitor of apoptosis as a novel prognostic marker in radically resected non-small cell lung cancer patients. , 2001, Clinical cancer research : an official journal of the American Association for Cancer Research.

[5]  N. Sonenberg,et al.  Internal ribosome initiation of translation and the control of cell death. , 2000, Trends in genetics : TIG.

[6]  David E. Misek,et al.  Discordant Protein and mRNA Expression in Lung Adenocarcinomas * , 2002, Molecular & Cellular Proteomics.

[7]  Michael Weller,et al.  Smac agonists sensitize for Apo2L/TRAIL- or anticancer drug-induced apoptosis and induce regression of malignant glioma in vivo , 2002, Nature Medicine.

[8]  Markus Rehm,et al.  Real-time single cell analysis of Smac/DIABLO release during apoptosis , 2003, The Journal of cell biology.

[9]  J P Pignon,et al.  Irinotecan plus fluorouracil and leucovorin for metastatic colorectal cancer. , 2001, The New England journal of medicine.

[10]  M. Rehm,et al.  Intracellular signaling dynamics during apoptosis execution in the presence or absence of X-linked-inhibitor-of-apoptosis-protein. , 2008, Biochimica et biophysica acta.

[11]  J. Minna,et al.  Smac mimetic increases chemotherapy response and improves survival in mice with pancreatic cancer. , 2010, Cancer research.

[12]  J. Adams.  The proteasome: a suitable antineoplastic target , 2004, Nature Reviews Cancer.

[13]  Ki-Seok Jang,et al.  Reduced Expression of Apaf-1 in Colorectal Adenocarcinoma Correlates with Tumor Progression and Aggressive Phenotype , 2007, Annals of Surgical Oncology.

[14]  Chen Kaiyun,et al.  Expression of X‐linked inhibitor of apoptosis protein in human colorectal cancer and its correlation with prognosis , 2009 .

[15]  James A. Wells,et al.  Small-Molecule Activators of a Proenzyme , 2009, Science.

[16]  J C Reed,et al.  Somatic Frameshift Mutations in the BAX Gene in Colon Cancers of the Microsatellite Mutator Phenotype , 1997, Science.

[17]  John Calvin Reed,et al.  Analysis of Apoptosis Protein Expression in Early-Stage Colorectal Cancer Suggests Opportunities for New Prognostic Biomarkers , 2005, Clinical Cancer Research.

[18]  Tom Fearn,et al.  Sensitivity and Specificity , 2009 .

[19]  S. Srinivasula,et al.  Cytochrome c and dATP-Dependent Formation of Apaf-1/Caspase-9 Complex Initiates an Apoptotic Protease Cascade , 1997, Cell.

[20]  C. Compton,et al.  The predictive value of apoptosis protease‐activating factor 1 in rectal tumors treated with preoperative, high‐dose‐rate brachytherapy , 2006, Cancer.

[21]  Matty P. Weijenberg,et al.  A genomic screen for genes upregulated by demethylation and histone deacetylase inhibition in human colorectal cancer , 2002, Nature Genetics.

[22]  Markus Rehm,et al.  APOPTO-CELL - a simulation tool and interactive database for analyzing cellular susceptibility to apoptosis , 2007, Bioinform..

[23]  Hao Liu,et al.  Expression of X-linked inhibitor of apoptosis protein in human colorectal cancer and its correlation with prognosis. , 2009, Journal of surgical oncology.

[24]  T. Fan,et al.  Pharmacokinetics and derivation of an anticancer dosing regimen for PAC-1, a preferential small molecule activator of procaspase-3, in healthy dogs , 2011, Investigational New Drugs.

[25]  Raimund Mannhold,et al.  IAP antagonists: promising candidates for cancer therapy. , 2010, Drug discovery today.

[26]  Seamus J. Martin,et al.  Analysis of the composition, assembly kinetics and activity of native Apaf‐1 apoptosomes , 2004, The EMBO journal.

[27]  G. Giaccone,et al.  Apoptosis: target of cancer therapy. , 2002, Clinical cancer research : an official journal of the American Association for Cancer Research.

[28]  J. Prehn,et al.  Apoptosis signaling proteins as prognostic biomarkers in colorectal cancer: a review. , 2009, Biochimica et biophysica acta.

[29]  S. Horvath,et al.  Expression of X-Linked Inhibitor of Apoptosis Protein Is a Strong Predictor of Human Prostate Cancer Recurrence , 2007, Clinical Cancer Research.

[30]  Jochen H M Prehn,et al.  Systems analysis of effector caspase activation and its control by X‐linked inhibitor of apoptosis protein , 2006, The EMBO journal.

[31]  G. Hannon,et al.  Control of translation and mRNA degradation by miRNAs and siRNAs. , 2006, Genes & development.

[32]  S. Loening,et al.  Different mRNA and protein expression of matrix metalloproteinases 2 and 9 and tissue inhibitor of metalloproteinases 1 in benign and malignant prostate tissue. , 2002, European urology.

[33]  W. Earnshaw,et al.  Induction of apoptosis by cancer chemotherapy. , 2000, Experimental cell research.

[34]  P. Hergenrother,et al.  Small-molecule activation of procaspase-3 to caspase-3 as a personalized anticancer strategy , 2006, Nature chemical biology.

[35]  P. Pasricha,et al.  Inhibition of apoptosis during development of colorectal cancer. , 1995, Cancer research.

[36]  D. Kuhn,et al.  Proteasome Inhibitors in Cancer Therapy: Lessons from the First Decade , 2008, Clinical Cancer Research.

[37]  C. Smith.  Diagnostic tests (1) – sensitivity and specificity , 2012, Phlebology.

[38]  Smac agonists sensitize for Apo 2 L / TRAILor anticancer drug-induced apoptosis and induce regression of malignant glioma in vivo , 2002, Nature Reviews Cancer.

[39]  W. Gerald,et al.  Inactivation of the apoptosis effector Apaf-1 in malignant melanoma , 2001, Nature.

[40]  E. Devilard,et al.  Caspase 7 downregulation as an immunohistochemical marker of colonic carcinoma. , 2001, Human pathology.

[41]  J. Herman,et al.  An inactivating mutation in HDAC2 leads to dysregulation of apoptosis mediated by APAF1. , 2008, Gastroenterology.

[42]  Seamus J. Martin,et al.  The Apoptosome Pathway to Caspase Activation in Primary Human Neutrophils Exhibits Dramatically Reduced Requirements for Cytochrome c , 2003, The Journal of experimental medicine.

[43]  D. Sargent,et al.  Prognostic impact of microsatellite instability and DNA ploidy in human colon carcinoma patients. , 2006, Gastroenterology.

[44]  A. Nesbakken,et al.  Tumour location is a prognostic factor for survival in colonic cancer patients , 2007, Colorectal disease : the official journal of the Association of Coloproctology of Great Britain and Ireland.

[45]  Guy S. Salvesen,et al.  X-linked IAP is a direct inhibitor of cell-death proteases , 1997, Nature.

[46]  Robert L Moritz,et al.  Identification of DIABLO, a Mammalian Protein that Promotes Apoptosis by Binding to and Antagonizing IAP Proteins , 2000, Cell.

[47]  T. Hickish,et al.  Oxaliplatin, fluorouracil, and leucovorin as adjuvant treatment for colon cancer. , 2004, The New England journal of medicine.

[48]  B. Vogelstein,et al.  A genetic model for colorectal tumorigenesis , 1990, Cell.

[49]  L. Terracciano,et al.  Role of APAF‐1, E‐cadherin and peritumoural lymphocytic infiltration in tumour budding in colorectal cancer , 2007, The Journal of pathology.

[50]  See “ An inactivating mutation in HDAC 2 leads to dysregulation of apoptosis mediated by APAF 1 , 2022 .